采用 PCR-RFLP 技术对来自中国不同地区猪源弓形虫的遗传特征进行分析。
Genetic characterization of Toxoplasma gondii from pigs from different localities in China by PCR-RFLP.
机构信息
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, PR China.
出版信息
Parasit Vectors. 2013 Aug 7;6:227. doi: 10.1186/1756-3305-6-227.
BACKGROUND
Toxoplasma gondii is a widely prevalent protozoan parasite that causes serious toxoplasmosis in humans and animals. The present study aimed to determine the genetic diversity of T. gondii isolates from pigs in Jiangxi, Sichuan, Guangdong Provinces and Chongqing Municipality in China using multilocous polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology.
METHODS
A total of 38 DNA samples were extracted from hilar lymph nodes of pigs with suspected toxoplasmosis, and were detected for the presence of T. gondii by semi-nested PCR of B1 gene. The positive DNA samples were typed at 11 genetic markers, including 10 nuclear loci, namely, SAG1, 5'-SAG2 and 3'-SAG2, alternative SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and an apicoplast locus Apico.
RESULTS
Twenty-five of the 38 DNA samples were T. gondii B1 gene positive. Complete genotyping data for all loci could be obtained for 17 of the 25 samples. Two genotypes were revealed (ToxoDB PCR-RFLP genotypes #9 and #3). Sixteen samples belong to genotype #9 which is the major lineage in mainland China and one sample belongs to genotype #3 which is Type II variant.
CONCLUSIONS
To our knowledge, this is the first report of genetic typing of T. gondii isolates from pigs in Jiangxi, Sichuan Province and Chongqing Municipality, and the first report of ToxoDB #3 T. gondii from pigs in China. These results have implications for the prevention and control of foodborne toxoplasmosis in humans.
背景
刚地弓形虫是一种广泛流行的原生动物寄生虫，可导致人类和动物发生严重的弓形体病。本研究旨在采用多位点聚合酶链反应-限制性片段长度多态性（PCR-RFLP）技术，检测来自中国江西、四川、广东和重庆的猪源弓形虫分离株的遗传多样性。
方法
从疑似弓形体病的猪的肺门淋巴结中提取 38 个 DNA 样本，通过 B1 基因半巢式 PCR 检测是否存在弓形虫。阳性 DNA 样本采用 11 个遗传标记（10 个核基因座，即 SAG1、5′-SAG2 和 3′-SAG2、替代 SAG2、SAG3、BTUB、GRA6、c22-8、c29-2、L358、PK1 和质体基因座 Apico）进行基因分型。
结果
38 个 DNA 样本中，有 25 个 B1 基因阳性。对 25 个样本中的 17 个样本获得了所有基因座的完整基因分型数据。揭示了两种基因型（ToxoDB PCR-RFLP 基因型#9 和#3）。16 个样本属于中国内地主要谱系的基因型#9，1 个样本属于基因型#3，即 II 型变体。
结论
据我们所知，这是首次报道江西、四川和重庆猪源弓形虫分离株的遗传分型，也是首次报道中国猪源 ToxoDB#3 弓形虫。这些结果对预防和控制人类食源性弓形体病具有重要意义。
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